Metabolic engineering of Yarrowia lipolytica for poly(ethylene terephthalate) degradation

Kosiorowska, Katarzyna E.; Biniarz, Piotr; Dobrowolski, Adam; Leluk, Karol; Mirończuk, Aleksandra M.

doi:10.1016/j.scitotenv.2022.154841

Cited by 21 publications

(5 citation statements)

References 35 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This is one of the available insertion sites described in the EasyCloneYALI System [15]. Although this system was optimized for Y. lipolytica GB20 strains, some of the sites have proven to be e ciently transcribed also in derivatives of strain A101 [19].…”

Section: Reporter System For Lamentous Strainsmentioning

confidence: 99%

Mutation in yl-HOG1 represses the filament-to-yeast transition in the dimorphic yeast Yarrowia lipolytica

Rzechonek

Szczepańczyk

Mirończuk

2023

Preprint

Self Cite

View full text Add to dashboard Cite

Background Yarrowia lipolytica is a dimorphic fungus, which switches from yeast to yeast-to-filament form in response to environmental conditions. For industrial purposes it is important to lock cells in the yeast or filamentous form depending on the fermentation process. yl-Hog1 kinase is a key component of the HOG signaling pathway, responsible for activating the osmotic stress response. Additionally, deletion of yl-Hog1 leads to increased filamentation in Yarrowia lipolytica, but causes significant sensitivity to osmotic stress induced by a high concentration of a carbon source. Results In this study, we tested the effect of point mutations on the function of yl-Hog1 protein kinase. The targets of modification were the phosphorylation sites (T171A-Y173A) and the active center (K49R). Introduction of the variant HOG1-49 into the hog1∆ strain partially improved growth under osmotic stress, but did not recover the yeast-like shape of the cells. The HOG1-171/173 variant was completely inactive, and its introduction further weakened the hog1∆ strains. To verify a genetic modification in filament form, we developed a new system based on green fluorescent protein (GFP) for easier screening of proper mutants. Conclusions These results provide new insights into the functions of yl-Hog1 protein in dimorphic transition and constitute a good starting point for further genetic modification of Y. lipolytica in filament form.

show abstract

Section: Reporter System For Lamentous Strainsmentioning

confidence: 99%

Mutation in yl-HOG1 represses the filament-to-yeast transition in the dimorphic yeast Yarrowia lipolytica

Rzechonek

Szczepańczyk

Mirończuk

2023

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…11 Structural similarity of TfCut2 (51%) with IsPETase is slightly more than that of LCC (49%), which makes TfCut2 a valid target for our study. 23 Moreover, the amino acid residues involved in the cleavage of ester bond in TfCut2 are the same as that of IsPETase. 12 Muller et al in 2005 rst reported the PET degradation by TfCut2 with a degradation rate of 2.5 nmol À1 min À1 cm À2 .…”

Section: Introductionmentioning

confidence: 99%

“…14,[20][21][22] Studies to improve the performance of these enzymes is indeed a valuable research area. [23][24][25] In the year 2022, a novel enzyme PHL7 was discovered by Zimmermann and co-workers, which shows promising activity for large scale degradation of PET. 26,27 In the same year, application of machine learning techniques are also used to design FAST-IsPETase, 28 which shows higher activity.…”

Section: Introductionmentioning

confidence: 99%

Cation–π and hydrophobic interaction controlled PET recognition in double mutated cutinase – identification of a novel binding subsite for better catalytic activity

James

2022

RSC Adv.

View full text Add to dashboard Cite

show abstract

“…Other examples of TPA upcycling include conversion into catechol, muconic acid, glycolic acid, and vanillic acid [ 26 , 27 ]. More recently, Pichia pastoris was shown to be a suitable platform for expression of PETase and Yarrowia lypolitica was shown to naturally degrade PET and metabolize EG and TPA [ 28 – 31 ].…”

Section: Introductionmentioning

confidence: 99%

Development of a yeast whole-cell biocatalyst for MHET conversion into terephthalic acid and ethylene glycol

et al. 2022

View full text Add to dashboard Cite

Background Over the 70 years since the introduction of plastic into everyday items, plastic waste has become an increasing problem. With over 360 million tonnes of plastics produced every year, solutions for plastic recycling and plastic waste reduction are sorely needed. Recently, multiple enzymes capable of degrading PET (polyethylene terephthalate) plastic have been identified and engineered. In particular, the enzymes PETase and MHETase from Ideonella sakaiensis depolymerize PET into the two building blocks used for its synthesis, ethylene glycol (EG) and terephthalic acid (TPA). Importantly, EG and TPA can be re-used for PET synthesis allowing complete and sustainable PET recycling. Results In this study we used Saccharomyces cerevisiae, a species utilized widely in bioindustrial fermentation processes, as a platform to develop a whole-cell catalyst expressing the MHETase enzyme, which converts monohydroxyethyl terephthalate (MHET) into TPA and EG. We assessed six expression architectures and identified those resulting in efficient MHETase expression on the yeast cell surface. We show that the MHETase whole-cell catalyst has activity comparable to recombinant MHETase purified from Escherichia coli. Finally, we demonstrate that surface displayed MHETase is active across a range of pHs, temperatures, and for at least 12 days at room temperature. Conclusions We demonstrate the feasibility of using S. cerevisiae as a platform for the expression and surface display of PET degrading enzymes and predict that the whole-cell catalyst will be a viable alternative to protein purification-based approaches for plastic degradation.

show abstract

Metabolic engineering of Yarrowia lipolytica for poly(ethylene terephthalate) degradation

Cited by 21 publications

References 35 publications

Mutation in yl-HOG1 represses the filament-to-yeast transition in the dimorphic yeast Yarrowia lipolytica

Mutation in yl-HOG1 represses the filament-to-yeast transition in the dimorphic yeast Yarrowia lipolytica

Cation–π and hydrophobic interaction controlled PET recognition in double mutated cutinase – identification of a novel binding subsite for better catalytic activity

Development of a yeast whole-cell biocatalyst for MHET conversion into terephthalic acid and ethylene glycol

Contact Info

Product

Resources

About